Method for processing blood samples in order to produce DNA complex patterns for diagnostic applications

ABSTRACT

A method for processing blood samples in order to produce DNA complex patterns for diagnostic applications. The method includes the aggregation and deposition of the DNA complex from the blood of a human being in order to form a unique pattern which can be used as a medical diagnostic tool to identify a change in the body caused by a specific physiological or pathological condition. The method identifies a change in the body by comparing the pattern of a person before a specific physiological or pathological condition to the pattern of a person after the condition. The specific condition causing the change is then determined by identifying and associating the unique pattern after the condition to the specific condition or disease based on comparative appearance. The present invention can be used as a diagnostic tool to aid in the determination of, among other things, the sex of a human fetus within a few days after conception, and the presence of a cancerous condition in the earliest stage of the disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application is a continuation of U.S. applicationSer. No. 09/764,783, filed Jan. 17, 2001.

BACKGROUND OF THE INVENTION

[0002] 1. Field of Invention

[0003] This invention relates to the field of medical diagnostictechniques, and more specifically to the processing and preparation ofblood samples to produce precipitated samples with unique andidentifiable patterns.

[0004] 2. Background of Invention

[0005] Traditional disease diagnosis normally begins with a physicalexamination of the patient's symptoms and a review of the patient'smedical and family histories. The diagnosis is usually confirmed as thepatient's disease progresses and additional symptoms appear. A properdiagnosis may often times require that laboratory tests be performed inorder to provide more detailed information about the patient'scondition. In this regard, there are numerous specific tests fordifferent diseases that have been developed and used in modern medicine.Some of the most common tests involve obtaining a blood sample from apatient and examining the sample in the laboratory for both the generaland specific diagnostic purposes. Since the circulation of blood is themost significant biochemical transporting and exchanging system in thehuman body, most illnesses will produce or induce certain changes in theblood's content or its properties. It does not matter whether the humanbody is exposed to a toxin released from a pathogen or from apathogen-induced immune response, or whether the body is under theinfluence of an endocrine system dysfunction or is experiencingnutritional or metabolic problems. All of the foregoing and any otherabnormal conditions have the potential to induce significant changes inthe blood. It is for this reason that blood samples have become the mostcommonly collected samples for the laboratory diagnosis and forensicevidence.

[0006] Macroscopically, the composition of blood is categorized into twomajor components: the cellular portion consisting of blood cells, andthe fluid portion called plasma. Normally, a blood sample will start tocoagulate after being extracted from a blood vessel. The coagulationprocess involves the aggregation of the blood cells to form a dark solidprecipitation. The remaining liquid portion of the blood is calledserum. If the blood sample is mixed with an anti-coagulant during orimmediately after collection, the blood will not coagulate and will stayin fluid form. If the anti-coagulated blood is centrifuged or kept in astationary state for a period of time, the blood cells will precipitate.The up-liquid portion of the anti-coagulated blood is the plasma.Depending on the test being conducted, the blood samples are normallycollected and used in these three forms: the whole blood, plasma orserum. Most laboratory tests, however, focus on the serum component ofcoagulated blood.

[0007] There are many prior art technologies for detecting changes inblood components and relating the changes as a diagnostic tool toindicate the presence of different diseases. For example, an increase ina patient's white blood cell count generally indicates an inflammationoccurring somewhere in the body. A low level hemoglobin number indicatesanemia. Without proper medication, diabetes patients will show anabnormal blood glucose level. Therefore, the changes of the bloodcomponents are not only resulting from the changes of blood systemitself but also can reflect many diseases on other parts of the body.Literally, hundreds of blood components have been identified and used toassist in diagnosing disease. There are still a significant portion ofcomponents and functions yet being identified. In this regard, themolecular changes of well-defined markers in the blood are readilydetected for many well defined and understood diseases. Most ofinfectious diseases that induce the immune responses are easilyidentified by the detection of antibodies in the blood. Unfortunately,for many less-well defined physiological illnesses, there are noapparent changes in the body, nor in the blood, until the relativelylate stages of the disease. Especially for many kinds of cancer, thereis no good marker available.

[0008] Genes were originally understood to be the basic molecular unitwhich controlled inheritance. After the development of the advancedbiochemical technology, the chemical structural unit of genes has beenrevealed to be nucleic acids. Except for a small number of RNA(ribonucleic acid) viruses, genes in all other living organisms areformed from DNA (deoxyribonucleic acid). DNA strands are extremely longand unbranched nucleic acid polymers that contain many different genesin each strand. In living cells, DNA strands fold tightly with certainproteins to form a compact nucleoprotein complex called chromatin.Normally, DNA strands in this form are not active. These sequesteredgenes can be activated, however, as a result of certain controlledmechanisms that are, in general, not well understood by today's science.Though some studies on the physical structure of the DNA double helixwere done in the early stages of DNA analysis by biochemists,biochemistry and molecular biology research done today on the structureand functionality of DNA are mostly confined to an analysis of DNA afterit has been almost completely isolated from its nucleoprotein complex.

[0009] During the later development stage of molecular biology, theidentification of Enhancers, a kind of gene expression regulator, andthe discovery of ribozymes have indicated that an analysis of thesecondary or tertiary structure of entire nucleic acid complex, beyondjust the linear sequence of the nucleic acid units, might have apotentially more important role in the analysis and understanding ofgene regulation. The gene rearrangement process in the immune systemalso has indicated that gene regulation is involved in changes insubstantial portions of DNA, instead of just an alteration of a fewnucleic acids that only affected a very small portion of entire DNAsequence. Unfortunately, however, due to the extremely long and thinstructure of DNA and its dense and compact nucleoprotein complex, it isextremely difficult to isolate the DNA without significantly alteringits nucleoprotein complex. Indeed, standard DNA extraction proceduresused in molecular biology research cause severe damage to the integrityof the DNA complex. These procedures strip off all of the DNA'sassociated proteins and related components in order to isolate the DNA.Unfortunately, all of the DNA complex's structure is demolished leavingDNA in an isolated and an approximately linear and uniform form. As aresult, it is not possible on a macroscopic level to ascertain anystructural difference between normal and altered DNA.

[0010] It would be greatly advantageous to overcome the foregoingproblem in order to enable researchers to distinguish normal DNA fromaltered DNA by providing them with a method for processing and preparingblood samples to produce precipitated samples with unique andidentifiable patterns so that they can compare the patterns associatedwith a normal DNA complex to the patterns associated with an abnormalDNA complex.

SUMMARY OF THE INVENTION

[0011] In order to overcome the technological barrier, the presentinvention provides a new and unique method for processing and preparingblood samples that isolates and preserves the structural integrity ofDNA and its associated nucleoprotein complex (“DNA complex”). The methodincludes the aggregation and deposition of the DNA complex from theblood of a human being in order to form a unique pattern which can beused as a medical diagnostic tool to identify a change in the bodycaused by a specific physiological or pathological condition. Theaggregation and deposition of the DNA complex from the blood results ina unique pattern. Changes in the body can be identified by comparingthese patterns derived before a specific physiological or pathologicalcondition to those derived after the condition. Further, the specificcondition causing the change can be determined by identifying andassociating the unique pattern after the condition to the specificcondition or disease based on comparative appearance. The presentinvention can be used as a diagnostic tool to aid in the determinationof, among other things, the sex of a human fetus within a few days afterconception, and the presence of a cancerous condition in the earlieststage of the disease. In these and other contexts, an importantadvantage of the present invention is the ability to detect changes inthe body during the early stages of a physiological or pathologicalcondition without damaging the integrity of the DNA complex.

DESCRIPTION OF THE DRAWINGS

[0012] The present invention is further described by reference to thefollowing drawings:

[0013]FIG. 1 is a diagrammatical illustration of the present method ofprocessing human blood samples.

[0014]FIG. 2A is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a femalefetus for six (6) weeks.

[0015]FIG. 2B is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a femalefetus for seven (7) weeks.

[0016]FIG. 2C is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a femalefetus for eight (8) weeks.

[0017]FIG. 2D is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a femalefetus for nine (9) weeks.

[0018]FIG. 2E is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a femalefetus for ten (10) weeks.

[0019]FIG. 2F is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying femaletwins.

[0020]FIG. 3A is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a malefetus for five (5) weeks.

[0021]FIG. 3B is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a malefetus for seven (7) weeks.

[0022]FIG. 3C is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a malefetus for nine (9) weeks.

[0023]FIG. 3D is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a malefetus for ten (10) weeks.

[0024]FIG. 3E is a diagram of the pattern formed using the presentinvention to process the blood from a pregnant woman carrying a malefetus for eleven (11) weeks.

[0025]FIG. 4 is a diagram of the pattern formed using the presentinvention to process the blood of a healthy woman.

[0026]FIG. 5 is a diagram of the pattern formed using the presentinvention to process the blood of a woman with breast cancer.

[0027]FIG. 6 is a diagram of the pattern formed using the presentinvention to process the blood of a woman with cervical cancer.

DETAILED DESCRIPTION OF THE INVENTION

[0028] The new and unique method of processing human blood samplesaccording to the present invention is illustrated diagrammatically inFIG. 1.

[0029] Initially, fresh human blood 10 is collected and placed in afirst centrifuge tube 20, where it is mixed with an anticoagulant toform an anticoagulated blood mixture 30. The centrifuge tube 20containing the anticoagulated blood mixture 30 is then placed in acentrifuge 40. Centrifugation causes the anticoagulated blood toseparate into a top portion, containing blood plasma 50, which isremoved and discarded, leaving behind blood cells 60 in centrifuge tube20.

[0030] In a second centrifuge tube 21, a blood cell solution is preparedin accordance with the following steps: first, approximately two (2)volumes of Tris-buffer is placed into the centrifuge tube 21; second,approximately one (1) volume of Tris-buffer saturated phenol, preparedby mixing re-distilled phenol with Tris-buffer, is added to centrifugetube 21; and lastly, approximately four (4) volumes of the blood cells60 are added to centrifuge tube 21. In each of the first two steps, theTris-buffer consists of 0.5 M Tris, 0.2 M EDTA, 0.6% NaCl, having a pHof between 10.3 and 10.4.

[0031] The blood cell solution is then mixed well in the centrifuge tube21 by strong vibration for approximately 30 seconds to form a firstblood cell mixture 80, and then the mixture 80 is placed in thecentrifuge 40 where the mixture is centrifuged at 11,000 rpm forapproximately 10 minutes to produce a first liquid phase 90 and firstblood cell debris 91.

[0032] Next, a phenol/chloroform solution 100, consisting ofapproximately one (1) volume of Tris-buffer saturated phenol, preparedas described above, and approximately one (1) volume of chloroform, isadded to the centrifuge tube 21 containing the first liquid phase 90 andfirst blood cell debris 91. The contents in centrifuge tube 21 are thenmixed well to produce a second blood cell mixture 110.

[0033] The centrifuge tube 21 containing the second blood cell mixture110 is placed in centrifuge 40 and centrifuged at approximately 11,000rpm for approximately 15 minutes to produce a second liquid phase 120and second blood cell debris 130. The tube 21 containing the secondliquid phase 120 and debris 130 is then placed in a beaker of ice 140for approximately fifteen (15) minutes.

[0034] A slide 160 is then prepared by placing an acid alcohol sample150 consisting of approximately 25 μl of freshly made 20% acid alcohol(i.e., ethanol containing 20% acetic acid) onto the top surface of theslide 160, and immediately adding a blood cell sample 170 consisting ofapproximately one (1) volume of the second liquid phase 120, which hasbeen cooled, onto the center portion of the top surface of acid alcoholsample 150. The slide 160 is then maintained on a stable surface inorder to allow acid alcohol sample 150 and blood cell sample 170 to dryon the slide at room temperature without any disturbance for 10 to 15minutes. During the drying phase, the DNA complex contained within bloodcell sample 170 aggregates in the acid alcohol sample 150 and deposits aunique pattern on slide 160.

[0035] The phenol does not strip off all of the nucleoprotein complex.Phenol by itself cannot strip off all of the complex (e.g., histoneprotein), albeit this can be done by treating the tissue with protease(e.g., protease K). The method of the present invention does not includethis step. Rather, the present invention relies, in part, upon the factthat the DNA structure changes as part of a rearrangement process whichtakes place in bone marrow and which has been shown to be critical tothe production of a vast variety of white blood cells with each varietycontaining a uniquely rearranged DNA structure. What is significant tothe present invention is that after the mature white blood cells enterthe blood stream, some of the cells produce specific clones as part ofan immunological response to the exposure to an antigen, such as aphysiological or pathological condition. If the physiological orpathological condition is significant (e.g., pregnancy or cancer), theimmunological response produces large numbers of specific clones whichare designed to protect the body from the antigen. The change in the DNAcomplex pattern of a healthy person after the person has been exposed toa physiological or pathological condition is most probably due to thissubstantial increase in the volume of specific white blood cell clonesas compared to the volume of all other white blood cells in the blood.Since the method of the present invention preserves the structuralintegrity of the DNA complex, the DNA complex tends to aggregate duringthe deposition phase in an orderly manner, rather than in a randommanner. This effect is enhanced by the change of surface tension duringthe acid alcohol drying process, a process similar to DNA combingeffect. It is this orderly aggregation process that produces the uniquepatterns.

[0036] The foregoing has been established through extensive experimentsprovided that the relative concentrations of blood cells, Tris-bufferdiluted phenol, Tris-buffer saturated phenol and chloroform aremaintained. Moreover, it has been found that the resulting DNA complexpatterns for any given blood sample from the same person with nophysiological or pathological changes will exhibit practically identicalshapes. However, physiological or pathological changes to that personimpart repeatable changes in the patterns. Consequently, visual analysisof the patterns under an optical microscope has revealed that thepatterns are useful as a diagnostic tool to assist in the determinationof whether a particular human being has been exposed to a specificphysiological or pathological condition. Changes in the patterns serveas an early indicator of the onset of a physiological or pathologicalcondition. Moreover, in the experimental contexts to be described thetype of change tends to indicate the type of physiological orpathological condition.

[0037] In one set of experiments, the inventors used the method of thepresent invention to produce DNA complex patterns of approximately 1,000pregnant women, and the pattern results are shown in FIGS. 2A through3E. FIGS. 2A through 3E are line drawings of selected representative DNAcomplex patterns derived from photographs taken through an opticalmicroscope during the above-described experimentation. Generally, it wasfound that the DNA complex patterns for any given blood sample from thesame person exhibited practically identical shapes. It was also foundthat the patterns were visually distinguishable in the earliest stagesof pregnancy.

[0038] In each instance, the following procedure was utilized. Afterobtaining whole blood from a woman, the blood was placed in a centrifugetube where it was mixed with an anti-coagulant to form ananti-coagulated blood mixture. The anti-coagulated blood is thencentrifuged and the plasma was removed, leaving the blood cells in thetube. In another centrifuge tube, a blood cell solution was prepared inaccordance with the following steps: first, two (2) volumes (5 μl) ofTris-buffer were placed into the centrifuge tube; second, one (1) volume(2.5 μl) of Tris-buffer saturated phenol, which was prepared by mixingre-distilled phenol with Tris-buffer, was added to the centrifuge tube;and lastly, four (4) volumes (10 μl) of the blood cells were added tothe centrifuge tube. In each of the first two steps, the Tris-bufferconsisted of 0.5 M Tris, 0.2 M EDTA, 0.6% NaCl, having a pH of between10.3 and 10.4. The blood cell solution was then mixed well to produce afirst blood cell mixture, and the mixture was centrifuged for ten (10)minutes at 11,000 rpm to produce a first liquid phase and a first bloodcell debris, which formed as a precipitate in the tube. Next, aphenol/chloroform solution was prepared by mixing 2.5 μl of Tris-buffersaturated phenol, prepared as described above, with 2.5 μl chloroform,and the phenol/chloroform solution was added to the centrifuge tubecontaining the first liquid phase and first blood cell debris, which wasmixed well to produce a second blood cell mixture.

[0039] The second blood cell mixture was then centrifuged for fifteen(15) minutes at 11,000 rpm to produce a second liquid phase and secondblood cell debris, which formed as a precipitate in the tube. Thecentrifuge tube containing the second liquid phase and second blood celldebris was then removed from the centrifuge and placed in a beaker ofice for fifteen (15) minutes.

[0040] A slide was prepared by placing an acid alcohol sample of 25 μlof freshly prepared 20% acid alcohol (i.e., ethanol containing 20%acetic acid) onto the top surface of the slide, and a blood cell sampleof 1.0 μl of the cooled second liquid phase was immediately added to thecenter of the top surface of the acid alcohol sample. The samples werethen allowed to dry on the slide at room temperature without anydisturbance for 10 to 15 minutes. After the samples had dried, the slidewas viewed under an optical microscope.

[0041] Microscopic analysis of each sample from the blood of thepregnant women participating in the experiment revealed that thepatterns that formed on the slide from women carrying a female fetuswere readily distinguishable from the patterns formed from womencarrying a male fetus. It was also discovered that the patterns weredistinguishable in the earliest stages of pregnancy. The patterndiagrams shown in FIGS. 2A through 2F were obtained by using the presentinvention, as described in the experimental procedure above, to processa blood sample obtained from each of six (6) different pregnant womencarrying a female fetus. The gestation period for the women carrying afemale fetus was between six (6) and ten (10) weeks. Each of thesepatterns exhibits a single and approximately circular or polygonal ringshape, with the exception of FIG. 2F which exhibits a double ring shapeformed from the blood of one of the women who was carrying twins.

[0042] By comparison, the pattern diagrams shown in FIGS. 3A through 3Ewere formed by using the present invention to process a blood sampleobtained from each of five (5) different women carrying a male fetus.The gestation period for the women carrying a male fetus was betweenfive (5) and eleven (11) weeks. Each of these patterns reveals either agenerally linear pattern, or a linear pattern in combination with one ormore elongated or collapsed ring patterns.

[0043] None of the patterns illustrated in FIGS. 3A through 3E, however,exhibits the circular or polygonal shapes exhibited by the patternsillustrated in FIGS. 2A through 2F. Thus, with regard to a pregnantwoman, if the pattern is circular or polygonal in shape, the fetus is afemale and if the pattern is linear or elongated in shape, the fetus isa male. Applicants' experiments with pregnant women and nonpregnantwomen also revealed that the DNA complex patterns for any given bloodsample from the same person exhibited practically identical shapes.

[0044] For purposes of the present invention, the contemplated patternrecognition is actually a pattern characterization. It is not aquantitative measurement. There is no numeric number or data thatresults as the characterization is a subjective “yes” or “no”.Consequently, conventional statistical analysis will not support orreject the method, and has no place. As an example, consider therestriction mapping of DNA samples in forensic analysis. What matters isthat the DNA fragment bands match, but the quantity has no meaning.Another example involves pathology inspection. In liver cirrhosis, forexample, all damaged liver cells are different but they all share somesimilar characters. The similarity can be visually identified bypathologists. However, there is no place for a statistical analysis onthese cells because the cell is irregular in shape, and quantitativeanalysis has no meaning for the disease. The question is whether thereis a cirrhosis or not, and not the percentage of livers with cirrhosis.The present method involves the same qualitative diagnosis, notquantitative.

[0045] In another set of experiments, the inventors used the method ofthe present invention to produce DNA complex patterns from the blood ofapproximately 300 healthy women and 300 healthy men. The inventors alsoused the method of the present invention to produce DNA complex patternsof over 100 women with cervical cancer, and over 50 with breast cancer.The healthy female patterns were compared to the patterns from womenwith breast and cervical cancer.

[0046]FIG. 4 diagrammatically illustrates a typical pattern formed froma healthy woman, and FIG. 5 and FIG. 6 illustrate the patterns of awoman with breast and cervical cancer, respectively.

[0047] The pattern from the healthy women, as shown in FIG. 4 displays agenerally linear, single continuous strand. Throughout most of thestrand's length it is smooth and of uniform width. In some healthypersons the strands are straight and in some they form a nearly circularshape, yet in all cases they retain a smooth and uniform width. Thesesmooth strand shapes are readily distinguishable from the irregularbeaded and branched strand shapes in women with cancer.

[0048] The pattern formed from the blood cells of a woman with breastcancer, as illustrated in FIG. 5, reveals a generally polygonal shapedstrand that is beaded and there is a substantial discontinuity on oneside of the polygonal shaped strand. In all breast cancer samples therewas a rough string with very obvious irregular width and always circularor near circular shape. Also, there were consistently two linked closedrings of unequal size. The shapes of the closed rings were irregular. Itcould be triangle, circular, polygonal, etc. The discontinuity appearsas a branching pattern over from the polygonal shaped strand.

[0049]FIG. 6 is an exemplary aggregated DNA pattern of a female withcervical cancer. In all such samples there was always a closed ring withpartially diffused string forming branches or mesh type distributionfilling in the central space of the closed ring. Also, the shape of therings was highly metamorphic (as was the breast cancer). This can beseen in FIG. 6 where the strand pattern that was formed appears to begenerally smooth, but its shape is irregular, being neither linear,circular, nor polygonal, and there is a discontinuity or loop on oneportion of the strand, and thinner strands form branches which extendfrom one end of the thicker strand to the other end and one thin strandextends across the interior of the irregularly shaped strand pattern.

[0050] It should be apparent that the foregoing analyses are qualitativepattern characterizations. They do not involve quantitativemeasurements. There is no numerical quantization of these patterns. Thecomparison is visual and the result is either “yes” or “no”, just as apathologist would diagnose a biopsy. Therefore, conventional statisticanalysis can not be applied. Still, the sample characteristics areconsistently the same.

[0051] Based upon the consistent results of these experiments, it isapparent that the non-invasive method at the present invention may beused by biomedical researchers and technologists to identifyphysiological changes in the body of a pregnant woman in the earlieststages of her pregnancy, and to determine whether the changes are theresult of carrying a male or female fetus. As described in detail above,applicants have identified two (2) predominate strand patterns from theblood of a pregnant woman which determine the sex of a fetus within afew weeks of conception. If the pattern is circular or polygonal inshape (composite FIG. 2), the fetus is a female and if the pattern islinear or elongated in shape (composite FIG. 3), the fetus is a male.Thus, the present method of processing human blood allows someoneskilled in the art to readily recognize changes in the predominatelysmooth strands by using an optical microscope to detect the presence orabsence of beads within a smooth strand, a loop within a strand and/orsignificant branching coming off of a smooth strand.

[0052] Similarly, the marked differences between the patterns of ahealthy women and women with cancer indicates that the present inventionmay also be used to identify pathological changes in a human being. Theinventor's experiments have correlated a beaded strand with somebranching to the presence of breast cancer (see FIG. 5) and havecorrelated a looped strand with some branching with cervical cancer (seeFIG. 6). These results are significant because they indicate that themethod can be used as a diagnostic screen to indicate whether a personis being subjected to a significant pathological condition, warrantingfurther medical evaluation and diagnostic techniques. In other words,the relatively simple blood test described in the specification can beused to tell a person whether or not he or she is healthy. If the testindicates the presence of disease, earlier detection would then bepossible using more expensive and invasive techniques like an MRI, a CATscan and a colonscopy. Naturally, a normal test result would provide ageneral sense of well being to the patient, possibly removing ageneralized fear that he or she has cancer or some other seriousdisease. Again, the method of the present invention preserves thestructural integrity of the DNA complex, the DNA complex tends toaggregate during the deposition phase in an orderly manner, rather thanin a random manner. It is this orderly aggregation process that producesthe unique patterns.

[0053] It should be further understood by those persons who are skilledin the art of biomedical research that the application of the presentinvention is not limited to its use in diagnosing the gender of a fetus,and in diagnosing breast and cervical cancer. Since the patterns whichare used to form the diagnosis are formed from the DNA complex of theblood cells, the existence of all physiological and pathologicalconditions are potentially detectable using the method of the invention.Although current biomedical technology is not able to completelydescribe all of the physical changes that take place within the DNAcomplex of the blood cells when it is exposed to physiological and/orpathological conditions, it is known that changes do occur in the DNAcomplex of the blood cells in response to such conditions. As a result,the present invention's ability to compare a DNA complex pattern of athe blood cells of a healthy person to the pattern of a DNA complexwhich has reacted or changed in some manner compels the conclusion thatthe method may be used to diagnose the existence of all known diseasesand to do so at an early stage of the disease.

[0054] Although the preferred method of the present invention involvesthe use of fresh blood as the source material to form the patterns, itwill be understood by biomedical researchers that other cellular sourcematerials may also be used to extract the DNA complex and used to formprecipitated patterns. Finally, it will also be apparent to thoseskilled in the art that the method of the present invention, in additionto being applicable to human beings, is equally applicable to all otheranimals or living cells. Accordingly, the disclosure of the presentinvention is intended to be illustrative, but not limiting, of the scopeof the invention.

We claim:
 1. A method of processing human blood samples to form a DNAcomplex strand pattern, comprising the steps of: a. mixing a sample ofblood containing plasma and blood cells with an anticoagulant to form ananti-coagulated blood mixture; b. centrifuging the anti-coagulated bloodmixture in order to separate the plasma from the blood cells; c.preparing a first blood cell mixture in accordance with the followingsteps: i. preparing approximately one (1) volume of Tris-buffer; ii.adding approximately a half (½) volume of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer, to theapproximately one (1) volume of Tris-buffer to produce a buffer dilutedphenol; and iii. adding approximately two (2) volumes of the blood cellsto the buffer diluted phenol; d. centrifuging the first blood cellmixture to form a first liquid phase and first blood cell debris; e.preparing a second blood cell mixture by mixing the centrifuged firstblood cell mixture and first blood cell debris with approximately a half(½) volume of chloroform and approximately a half (½) volume ofTris-buffer saturated phenol, prepared by mixing re-distilled phenolwith Tris-buffer; f. centrifuging the second blood cell mixture to forma second liquid phase and second blood cell debris; g. cooling thesecond liquid phase and second blood cell debris thereby causing thestructural components of the DNA complex within the second liquid phaseto aggregate; h. placing an acid alcohol sample consisting ofapproximately twelve and a half (12½) volumes of freshly made 20% acidalcohol on a slide; and i. adding a blood cell sample consisting ofapproximately one fifth (⅕) volume of the cooled second liquid phaseonto the center of the top surface of the acid alcohol sample andallowing both samples to dry at room temperature without anydisturbance, whereby an aggregate of the DNA complex deposits a strandpattern on the slide.
 2. A method of processing human blood samples toform a DNA complex strand pattern, comprising the steps of: a. mixing asample of blood containing plasma and blood cells with an anticoagulantto form an anti-coagulated blood mixture; b. centrifuging theanti-coagulated blood mixture in order to separate the plasma from theblood cells; c. preparing a first blood cell mixture in accordance withthe following steps: i. preparing approximately 5 ml of Tris-buffer; ii.adding approximately 2.5 ml of Tris-buffer saturated phenol, prepared bymixing re-distilled phenol with Tris-buffer, to the approximately 5.0 mlof Tris-buffer to produce a buffer diluted phenol; and iii. addingapproximately 10 ml of the blood cells to the buffer diluted phenol; d.centrifuging the first blood cell mixture to form a first liquid phaseand first blood cell debris; e. preparing a second blood cell mixture bymixing the centrifuged first blood cell mixture and first blood celldebris with approximately 2.5 ml of chloroform and approximately 2.5 mlof Tris-buffer saturated phenol, prepared by mixing re-distilled phenolwith Tris-buffer; f. centrifuging the second blood cell mixture to forma second liquid phase and second blood cell debris; g. cooling thesecond liquid phase and second blood cell debris thereby causing thestructural components of the DNA complex within the second liquid phaseto aggregate; h. placing an acid alcohol sample consisting ofapproximately 25 ml of freshly made 20% acid alcohol on a slide; i.adding a blood cell sample consisting of approximately 1.0 ml of thecooled second liquid phase onto the center of the top surface of theacid alcohol sample and allowing both samples to dry at room temperaturewithout any disturbance, whereby an aggregate of the DNA complexdeposits a strand pattern on the slide.
 3. A method of determining thesex of a fetus comprising: a. mixing a sample of blood containing plasmaand blood cells with an anticoagulant to form an anti-coagulated bloodmixture; b. centrifuging the anti-coagulated blood mixture in order toseparate the plasma from the blood cells; c. preparing a first bloodcell mixture in accordance with the following steps: i. preparingapproximately one (1) volume of Tris-buffer; ii. adding approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer, to the approximately one (1)volume of Tris-buffer to produce a buffer diluted phenol; and iii.adding approximately two (2) volumes of the blood cells to the bufferdiluted phenol; iv. mixing the first blood cell mixture; d. centrifugingthe first blood cell mixture to form a first liquid phase and firstblood cell debris; e. preparing a second blood cell mixture by mixingthe centrifuged first blood cell mixture and first blood cell debriswith approximately a half (½) volume of chloroform and approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately twelve and a half (12½) volumes of freshly made 20%acid alcohol on a slide; and i. adding a blood cell sample consisting ofapproximately one fifth (⅕) volume of the cooled second liquid phaseonto the center of the top surface of the acid alcohol sample andallowing both samples to dry at room temperature without anydisturbance, whereby an aggregate of the DNA complex deposits a strandpattern on the slide; and J. determining that the sex of the fetus isfemale if the shape of the strand pattern is approximately circular orpolygonal, or that the sex of the fetus is male if the shape of thestrand pattern is generally linear or generally linear in combinationwith at least one elongated ring.
 4. A method of determining the sex ofa fetus comprising: a. mixing a sample of blood containing plasma andblood cells with an anticoagulant to form an anti-coagulated bloodmixture; b. centrifuging the anti-coagulated blood mixture in order toseparate the plasma from the blood cells; c. preparing a first bloodcell mixture in accordance with the following steps: i. preparingapproximately 5 μl of Tris-buffer; ii. adding approximately 2.5 μl ofTris-buffer saturated phenol, prepared by mixing re-distilled phenolwith Tris-buffer, to the approximately 5.0 μl of Tris-buffer to producea buffer diluted phenol; and iii. adding approximately 10 μl of theblood cells to the buffer diluted phenol; iv. mixing the first bloodcell mixture; d. centrifuging the first blood cell mixture to form afirst liquid phase and first blood cell debris; e. preparing a secondblood cell mixture by mixing the centrifuged first blood cell mixtureand first blood cell debris with approximately 2.5 μl of chloroform andapproximately 2.5 μl of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately 25 μl of freshly made 20% acid alcohol on a slide; i.adding a blood cell sample consisting of approximately 1.0 μl of thecooled second liquid phase onto the center of the top surface of theacid alcohol sample and allowing both samples to dry at room temperaturewithout any disturbance, whereby an aggregate of the DNA complexdeposits a strand pattern on the slide; and j. determining that the sexof the fetus is female if the shape of the strand pattern isapproximately circular or polygonal, or that the sex of the fetus ismale if the shape of the strand pattern is generally linear or generallylinear in combination with at least one elongated ring.
 5. The method ofclaim 3 or 4 in which the Tris-buffer consists of 0.5 M Tris, 0.2 MEDTA, 0.6% NaCl, having a pH of between 10.3 and 10.4.
 6. The method ofclaim 3 or 4 in which the step of centrifuging the first blood cellmixture is performed for approximately ten (10) minutes at 11,000 rpm.7. The method of claim 3 or 4 in which the step of centrifuging thesecond blood cell mixture is performed for approximately fifteen (15)minutes at 11,000 rpm.
 8. The method of claim 3 or 4 in which the stepof cooling the second liquid phase is performed by placing the secondliquid phase on ice for approximately fifteen (15) minutes.
 9. A methodof detecting a change in the body of a human being caused by apathological condition, comprising: a. mixing a sample of blood from ahuman donor with an anticoagulant to form an anti-coagulated bloodmixture; b. centrifuging the anti-coagulated blood mixture in order toseparate the plasma from the blood cells; c. preparing a first bloodcell mixture in accordance with the following steps: i. preparingapproximately one (1) volume of Tris-buffer; ii. adding approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer, to the approximately one (1)volume of Tris-buffer to produce a buffer diluted phenol; and iii.adding approximately two (2) volumes of the blood cells to the bufferdiluted phenol; iv. mixing the first blood cell mixture; d. centrifugingthe first blood cell mixture to form a first liquid phase and firstblood cell debris; e. preparing a second blood cell mixture by mixingthe centrifuged first blood cell mixture and first blood cell debriswith approximately a half (½) volume of chloroform and approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately twelve and a half (12½) volumes of freshly made 20%acid alcohol on a slide; i. adding a blood cell sample consisting ofapproximately one fifth (⅕) volume of the cooled second liquid phaseonto the center of the top surface of the acid alcohol sample andallowing both samples to dry at room temperature without anydisturbance, whereby an aggregate of the DNA complex deposits a strandpattern on the slide; and j. using the strand pattern to detect a changein the body of the human donor.
 10. A method of detecting a change inthe body of a human being caused by a pathological condition,comprising: a. mixing a sample of the human blood with an anticoagulantto form an anti-coagulated blood mixture; b. centrifuging theanti-coagulated blood mixture in order to separate the plasma from theblood cells; c. preparing a first blood cell mixture in accordance withthe following steps: i. preparing approximately 5 μl of Tris-buffer; ii.adding approximately 2.5 μl of Tris-buffer saturated phenol, prepared bymixing re-distilled phenol with Tris-buffer, to the approximately 5.0 μlof Tris-buffer to produce a buffer diluted phenol; and iii. addingapproximately 10 μl of the blood cells to the buffer diluted phenol; iv.mixing the first blood cell mixture; d. centrifuging the first bloodcell mixture to form a first liquid phase and first blood cell debris;e. preparing a second blood cell mixture by mixing the centrifuged firstblood cell mixture and first blood cell debris with approximately 2.5 mlof chloroform and approximately 2.5 ml of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer; f. centrifugingthe second blood cell mixture to form a second liquid phase and secondblood cell debris; g. cooling the second liquid phase and second bloodcell debris thereby causing the structural components of the DNA complexwithin the second liquid phase to aggregate; h. placing an acid alcoholsample consisting of approximately 25 μl of freshly made 20% acidalcohol on a slide; and i. adding a blood cell sample consisting ofapproximately 1.0 μl of the cooled second liquid phase onto the centerof the top surface of the acid alcohol sample and allowing both samplesto dry at room temperature without any disturbance, whereby an aggregateof the DNA complex deposits a strand pattern on the slide; and j. usingthe strand pattern to detect a change in the body of the human donor.11. A method of detecting a change in the body of a human being causedby a pathological condition, comprising: a. mixing a sample of bloodfrom a human donor with an anticoagulant to form an anti-coagulatedblood mixture; b. centrifuging the anti-coagulated blood mixture inorder to separate the plasma from the blood cells; c. preparing a firstblood cell mixture in accordance with the following steps: i. preparingapproximately one (1) volume of Tris-buffer; ii. adding approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer, to the approximately one (1)volume of Tris-buffer to produce a buffer diluted phenol; and iii.adding approximately two (2) volumes of the blood cells to the bufferdiluted phenol; iv. mixing the first blood cell mixture; d. centrifugingthe first blood cell mixture to form a first liquid phase and firstblood cell debris; e. preparing a second blood cell mixture by mixingthe centrifuged first blood cell mixture and first blood cell debriswith approximately a half (½) volume of chloroform and approximately ahalf (½) volume of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately twelve and a half (12½) volumes of freshly made 20%acid alcohol on a slide; i. adding a blood cell sample consisting ofapproximately one fifth (⅕) volume of the cooled second liquid phaseonto the center of the top surface of the acid alcohol sample andallowing both samples to dry at room temperature without anydisturbance, whereby an aggregate of the DNA complex deposits a strandpattern on the slide; and j. detecting a change in the body of the humandonor if the strand pattern comprises a strand which is not smooththroughout most of the strand's length.
 12. A method of detecting achange in the body of a human being caused by a pathological condition,comprising: a. mixing a sample of blood from a human donor with ananticoagulant to form an anti-coagulated blood mixture; b. centrifugingthe anti-coagulated blood mixture in order to separate the plasma fromthe blood cells; c. preparing a first blood cell mixture in accordancewith the following steps: i. preparing approximately one (1) volume ofTris-buffer; ii. adding approximately a half (½) volume of Tris-buffersaturated phenol, prepared by mixing re-distilled phenol withTris-buffer, to the approximately one (1) volume of Tris-buffer toproduce a buffer diluted phenol; and iii. adding approximately two (2)volumes of the blood cells to the buffer diluted phenol; iv. mixing thefirst blood cell mixture; d. centrifuging the first blood cell mixtureto form a first liquid phase and first blood cell debris; e. preparing asecond blood cell mixture by mixing the centrifuged first blood cellmixture and first blood cell debris with approximately a half (½) volumeof chloroform and approximately a half (½) volume of Tris-buffersaturated phenol, prepared by mixing re-distilled phenol withTris-buffer; f. centrifuging the second blood cell mixture to form asecond liquid phase and second blood cell debris; g. cooling the secondliquid phase and second blood cell debris thereby causing the structuralcomponents of the DNA complex within the second liquid phase toaggregate; h. placing an acid alcohol sample consisting of approximatelytwelve and a half (12½) volumes of freshly made 20% acid alcohol on aslide; i. adding a blood cell sample consisting of approximately onefifth (⅕) volume of the cooled second liquid phase onto the center ofthe top surface of the acid alcohol sample and allowing both samples todry at room temperature without any disturbance, whereby an aggregate ofthe DNA complex deposits a strand pattern on the slide; and j. detectinga change in the body of the human donor if the strand pattern comprisesa strand which has a plurality of beads and a substantial discontinuitywith associated branching.
 13. A method of detecting a change in thebody of a human being caused by a pathological condition, comprising: a.mixing a sample of blood from a human donor with an anticoagulant toform an anti-coagulated blood mixture; b. centrifuging theanti-coagulated blood mixture in order to separate the plasma from theblood cells; c. preparing a first blood cell mixture in accordance withthe following steps: i. preparing approximately one (1) volume ofTris-buffer; ii. adding approximately a half (½) volume of Tris-buffersaturated phenol, prepared by mixing re-distilled phenol withTris-buffer, to the approximately one (1) volume of Tris-buffer toproduce a buffer diluted phenol; and iii. adding approximately two (2)volumes of the blood cells to the buffer diluted phenol; iv. mixing thefirst blood cell mixture; d. centrifuging the first blood cell mixtureto form a first liquid phase and first blood cell debris; e. preparing asecond blood cell mixture by mixing the centrifuged first blood cellmixture and first blood cell debris with approximately a half (½) volumeof chloroform and approximately a half (½) volume of Tris-buffersaturated phenol, prepared by mixing re-distilled phenol withTris-buffer; f. centrifuging the second blood cell mixture to form asecond liquid phase and second blood cell debris; g. cooling the secondliquid phase and second blood cell debris thereby causing the structuralcomponents of the DNA complex within the second liquid phase toaggregate; h. placing an acid alcohol sample consisting of approximatelytwelve and a half (12½) volumes of freshly made 20% acid alcohol on aslide; i. adding a blood cell sample consisting of approximately onefifth (⅕) volume of the cooled second liquid phase onto the center ofthe top surface of the acid alcohol sample and allowing both samples todry at room temperature without any disturbance, whereby an aggregate ofthe DNA complex deposits a strand pattern on the slide; and j. detectinga change in the body of the human donor if the strand pattern comprisesa strand which has a looped portion and a substantial discontinuity withassociated branching.
 14. A method of detecting a change in the body ofa human being caused by a pathological condition, comprising: a. mixinga sample of the human blood with an anticoagulant to form ananti-coagulated blood mixture; b. centrifuging the anti-coagulated bloodmixture in order to separate the plasma from the blood cells; c.preparing a first blood cell mixture in accordance with the followingsteps: i. preparing approximately 5 μl of Tris-buffer; ii. addingapproximately 2.5 μl of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer, to the approximately 5.0 μl ofTris-buffer to produce a buffer diluted phenol; and iii. addingapproximately 10 μl of the blood cells to the buffer diluted phenol; iv.mixing the first blood cell mixture; d. centrifuging the first bloodcell mixture to form a first liquid phase and first blood cell debris;e. preparing a second blood cell mixture by mixing the centrifuged firstblood cell mixture and first blood cell debris with approximately 2.5 mlof chloroform and approximately 2.5 ml of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer; f. centrifugingthe second blood cell mixture to form a second liquid phase and secondblood cell debris; g. cooling the second liquid phase and second bloodcell debris thereby causing the structural components of the DNA complexwithin the second liquid phase to aggregate; h. placing an acid alcoholsample consisting of approximately 25 μl of freshly made 20% acidalcohol on a slide; and i. adding a blood cell sample consisting ofapproximately 1.0 μl of the cooled second liquid phase onto the centerof the top surface of the acid alcohol sample and allowing both samplesto dry at room temperature without any disturbance, whereby an aggregateof the DNA complex deposits a strand pattern on the slide; and j.detecting a change in the body of the human donor if the strand patterncomprises a strand which is not smooth throughout most of the strand'slength.
 15. A method of detecting a change in the body of a human beingcaused by a pathological condition, comprising: a. mixing a sample ofthe human blood with an anticoagulant to form an anti-coagulated bloodmixture; b. centrifuging the anti-coagulated blood mixture in order toseparate the plasma from the blood cells; c. preparing a first bloodcell mixture in accordance with the following steps: i. preparingapproximately 5 μl of Tris-buffer; ii. adding approximately 2.5 μl ofTris-buffer saturated phenol, prepared by mixing re-distilled phenolwith Tris-buffer, to the approximately 5.0 μl of Tris-buffer to producea buffer diluted phenol; and iii. adding approximately 10 μl of theblood cells to the buffer diluted phenol; iv. mixing the first bloodcell mixture; d. centrifuging the first blood cell mixture to form afirst liquid phase and first blood cell debris; e. preparing a secondblood cell mixture by mixing the centrifuged first blood cell is mixtureand first blood cell debris with approximately 2.5 ml of chloroform andapproximately 2.5 ml of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately 25 μl of freshly made 20% acid alcohol on a slide; andi. adding a blood cell sample consisting of approximately 1.0 μl of thecooled second liquid phase onto the center of the top surface of theacid alcohol sample and allowing both samples to dry at room temperaturewithout any disturbance, whereby an aggregate of the DNA complexdeposits a strand pattern on the slide; and j. detecting a change in thebody of the human donor if the strand pattern comprises a strand whichhas a plurality of beads and a substantial discontinuity with associatedbranching.
 16. A method of detecting a change in the body of a humanbeing caused by a pathological condition, comprising: a. mixing a sampleof the human blood with an anticoagulant to form an anti-coagulatedblood mixture; b. centrifuging the anti-coagulated blood mixture inorder to separate the plasma from the blood cells; c. preparing a firstblood cell mixture in accordance with the following steps: i. preparingapproximately 5 μl of Tris-buffer; ii. adding approximately 2.5 μl ofTris-buffer saturated phenol, prepared by mixing re-distilled phenolwith Tris-buffer, to the approximately 5.0 μl of Tris-buffer to producea buffer diluted phenol; and iii. adding approximately 10 μl of theblood cells to the buffer diluted phenol; iv. mixing the first bloodcell mixture; d. centrifuging the first blood cell mixture to form afirst liquid phase and first blood cell debris; e. preparing a secondblood cell mixture by mixing the centrifuged first blood cell mixtureand first blood cell debris with approximately 2.5 ml of chloroform andapproximately 2.5 ml of Tris-buffer saturated phenol, prepared by mixingre-distilled phenol with Tris-buffer; f. centrifuging the second bloodcell mixture to form a second liquid phase and second blood cell debris;g. cooling the second liquid phase and second blood cell debris therebycausing the structural components of the DNA complex within the secondliquid phase to aggregate; h. placing an acid alcohol sample consistingof approximately 25 μl of freshly made 20% acid alcohol on a slide; andi. adding a blood cell sample consisting of approximately 1.0 μl of thecooled second liquid phase onto the center of the top surface of theacid alcohol sample and allowing both samples to dry at room temperaturewithout any disturbance, whereby an aggregate of the DNA complexdeposits a strand pattern on the slide; and j. detecting a change in thebody of the human donor if the strand pattern comprises a strand whichhas a looped portion and a substantial discontinuity with associatedbranching.
 17. A method of detecting a change in the body of a humanbeing caused by a pathological condition, comprising: a. mixing a sampleof blood from a human donor with an anticoagulant to form ananti-coagulated blood mixture; b. centrifuging the anti-coagulated bloodmixture in order to separate the plasma from the blood cells; c.preparing a first blood cell mixture in accordance with the followingsteps: i. preparing approximately one (1) volume of Tris-buffer; ii.adding approximately a half (½) volume of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer, to theapproximately one (1) volume of Tris-buffer to produce a buffer dilutedphenol; and iii. adding approximately two (2) volumes of the blood cellsto the buffer diluted phenol; iv. mixing the first blood cell mixture;d. centrifuging the first blood cell mixture to form a first liquidphase and first blood cell debris; e. preparing a second blood cellmixture by mixing the centrifuged first blood cell mixture and firstblood cell debris with approximately a half (½) volume of chloroform andapproximately a half (½) volume of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer; f. centrifugingthe second blood cell mixture to form a second liquid phase and secondblood cell debris; g. cooling the second liquid phase and second bloodcell debris thereby causing the structural components of the DNA complexwithin the second liquid phase to aggregate; h. placing an acid alcoholsample consisting of approximately twelve and a half (12½) volumes offreshly made 20% acid alcohol on a slide; i. adding a blood cell sampleconsisting of approximately one fifth (⅕) volume of the cooled secondliquid phase onto the center of the top surface of the acid alcoholsample and allowing both samples to dry at room temperature without anydisturbance, whereby an aggregate of the DNA complex deposits a strandpattern on the slide; and j. using the strand pattern to detect a changein the body of the human donor.
 18. A method of detecting a change inthe body of a human being caused by a pathological condition,comprising: a. mixing a sample of the human blood with an anticoagulantto form an anti-coagulated blood mixture; b. centrifuging theanti-coagulated blood mixture in order to separate the plasma from theblood cells; c. preparing a first blood cell mixture in accordance withthe following steps: i. preparing approximately 5 μl of Tris-buffer; ii.adding approximately 2.5 μl of Tris-buffer saturated phenol, prepared bymixing re-distilled phenol with Tris-buffer, to the approximately 5.0 μlof Tris-buffer to produce a buffer diluted phenol; and iii. addingapproximately 10 μl of the blood cells to the buffer diluted phenol; iv.mixing the first blood cell mixture; d. centrifuging the first bloodcell mixture to form a first liquid phase and first blood cell debris;e. preparing a second blood cell mixture by mixing the centrifuged firstblood cell mixture and first blood cell debris with approximately 2.5 mlof chloroform and approximately 2.5 ml of Tris-buffer saturated phenol,prepared by mixing re-distilled phenol with Tris-buffer; f. centrifugingthe second blood cell mixture to form a second liquid phase and secondblood cell debris; g. cooling the second liquid phase and second bloodcell debris thereby causing the structural components of the DNA complexwithin the second liquid phase to aggregate; h. placing an acid alcoholsample consisting of approximately 25 μl of freshly made 20% acidalcohol on a slide; and i. adding a blood cell sample consisting ofapproximately 1.0 μl of the cooled second liquid phase onto the centerof the top surface of the acid alcohol sample and allowing both samplesto dry at room temperature without any disturbance, whereby an aggregateof the DNA complex deposits a strand pattern on the slide; and j. usingthe strand pattern to detect a change in the body of the human donor.